Al-Anbar J. Vet. Sci., Vol.: 5 No. (2), 2012 ISSN:

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1 Prevalence of Methicillin Resistance Staphylococcus aureus in Cattle and She-camels Milk at Al-Qadisyia Province M. H. Abdulkadhim College of Agriculture\ University of Al-Qadisiya Abstract This study is conducted to investigate the methicillin resistant Staphylococcus aureus (MRSA) in the cattle and she-camels milk. In which the number of examined milk samples in cattle and she-camels was (400, 360) with percentage of S. aureus (19.4%, 13.4%) respectively. Actually the subclinical mastitis widely distributed (59%, 43.3%) with the methicillin resistant isolates (39.1%, 0%) respectively in the both species. Furthermore the antibiotics sensitivity test reveals high susceptibility of this pathogens to cefoxitin and streptomycin and to several antibiotic types in the she-camels isolates. انتشار المكورات العنقودية المقاومة للمثيسيلين في حليب الابقار والنوق في محافظة القادسية محمد حمزة عبد الكاظم كلية الزراعة/ جامعة القادسية الخلاصة هدفت الدراسة الحالية للكشف عن جراثيم المكورات العنقودية المقاومة للمثيسلين في حليب الابقار والنوق اذ تم فحص ( ) عينه لكلا النوعين قد شخصت نسبة التهاب الضرع فيهما ( )% وعلى التوالي وقد بلغت نسبة الاصابة بتلك الجراثيم ( )% بينما بلغت نسبة الجراثيم المقاومة للمثيسيلين( )% وعلى التوالي. لقد اظهر فحص حساسية المضادات الحياتية فعالية المضاد سيفوكستين وستربتومايسين في تثبيط العزلات الجرثومية لحليب الابقار بينما كانت العزلات الجرثومية لحليب النوق حساسة لجميع المضادات الحياتية. Introduction Mastitis is considered one of the most important dairy cattle diseases and antibiotics use (1). The S. aureus is well known as causative agent of cattle and she-camels mastitis and characterized by acute, chronic, contagious and subclinical in nature (2, 3). Treatment of infections caused by S. aureus has become more difficult due to development of antimicrobial resistance, recently the MRSA widely distributed in all world with the risk of resistant to all beta-lactam antibiotics and other groups of antimicrobials therefore the therapeutic choices are limited significantly (4). The occurrence of MRSA in herds will develop a delay in the treatment of animal and human diseases with spreading in environment, therefore it has great zoonotic importance (5, 6, 7), therefore the aim of this study is to investigate the occurrence of MRSA in cattle and she-camels milk by using routine techniques, new chromogenic media and assess their antibiotics susceptibility to develop effective control strategies for this pathogens. Materials and Methods This study includes collection of 400, 360 milk samples of cattle and she-camels taken from different farms in Al-Qadisyia province under aseptic condition, the California mastitis test composed of Alkyl Aryl sulfonate 3%, sodium hydroxide 1.5% and bromocresol purple as an indicator is done on milk samples to identify the subclinical cases from the health ones while the clinical cases had been excluded from this study, the positive 63

2 isolates are examined in laboratory of veterinary hospital in Al-Qadisyia by inoculation and culturing at 37º for 24h on the CHROMagar Staph medium (CAS) then S. aureus specific colony is streaked in quadrant manner on MRSA CHROMagar medium at same laboratory conditions, its composition in g/l (agar 15; peptone and yeast extract 40; salts 25; chromogenic mix 2.5 aren't described by the manufacturer; PH: 6.9) (CHROMagar, France), all these was done after sub culturing and identification by conventional tests which includes (oxidase, coagulase, catalase, mannitol, sucrose, D. trehalose, gelatinase, urease, phosphatase, nitrate reductase), the MRSA isolates found are evaluated with antibiotics sensitivity test according the Kirby-Bauer Method by using four antibiotics which ordinarily used for treatment of cattle, she-camels and human diseases, these antibiotics are: oxacillin (Ox-1), cefoxitin (Fox30), penicillin G (Gp-10) and streptomycin (S10), its potency are (1µg, 30µg, 10 unit and 10µcg) respectively and the standard zone of inhibition are ( 13, 22, 29, 15 mm) respectively (Difico, USA) (8). Results The California mastitis test reveals in cattle and she-camels 236, 156 infected samples from total of 400, 360 milk samples respectively, these results show a significant differences (P<0.01) in the subclinical mastitis between the two types, but the infection percentage with S. aureus pathogens isn't significant (P>0.05) (Table 1). The S. aureus observed on (CAS, MRSA) culturing mauve and rose in color. The percentage of methicillin resistance S. aureus is 39.1% in mastitic cattle while there is no resistance compared with she-camels (P<0.01), furthermore the conventional microbiological tests confirmed these isolates and biochemically it was negative to oxidase but positive to coagulase, catalase, mannitol, sucrose, D. trehalose, gelatinase, urease, phosphatase and nitrate reductase. Additionally, it has characteristic growing colonies in blood agar (βhemolysis), mannitol salt agar, DNase and SM110 media. The antibiotics sensitivity test detect high susceptibility to cefoxitin and streptomycin (80.4, 78.2)% in cattle respectively, while in she-camels there is high susceptibility to the all antibiotics without any resistance to oxacillin and penicillin G (Table 2). Table (1) Show the percentage rate of infection with S. aureus No. of examined The infection Subclinical mastitis quarters S. aureus methicillin resistance + - % + - % + - % Cattle ** ** She-camels Chi 2 values X 2 tab. X 2 cal. X 2 tab. X 2 cal. X 2 tab. X 2 cal Table (2) Show the susceptibility of S. aureus to antibiotics Antibiotics cattle 64 She-camels R I S % R I S % Ox Fox Gp S Discussion The isolation of S. aureus pathogens from raw cattle and she-camels milk has been described from different parts of the world namely Saudi Arabia (9), Jordan (10, 11), Egypt (12) and Morocco (13). In the present study the incidence of subclinical mastitis in cattle and she-camels is relatively comparative with those reported by (14, 15) (80, 36)% respectively these may be due to differences of geographical regions. The prevalence of

3 subclinical mastitis in cattle and she-camels differs significantly (P<0.01), these results may be due to feature of the anatomical structure of the she-camels udder teats, glands cisterns which are separated entirely with very narrow twin duct streak canals. Furthermore the udder didn't touch the earth even the recumbent of the animal this act as an important mechanical prevention to infection (16), However, many researchers demonstrated that shecamels milk contains inhibitors to pathogenic bacteria these includes: Lysozyme, immunoglobulins, lactoferrin and lactoperoxidase, these proteins involved with higher concentration in she-camels milk than in cattle milk (17, 18, 19, 20, 21), As well as there is another difference related to nature environments between cattle and camels which characterized by less contaminators than cattle environment (22). The percentage of MRSA isolated from cattle higher than 10.2% which reported by (23) these differences may be due to variation of the geographical regions. The causes of antibiotic resistance is explained by several authors reported numerous resistance methods such as: antibiotic-resistant genes, mutation, clonal evolution and plasmid transfer, target site alteration of ribosome, metabolic pathway alteration, efflux pumps and enzymatic cleavage of antibiotics (23, 24), recently (25) has discussed the causes of these resistance as a results of a protein (penicillin-binding protein 2a[PBP2a]). The results have also revealed high susceptibility and absence of antibiotics resistance in S. aureus isolated from she-camels milk, actually recent studies have showed absence of meca gene responsible for MRSA and rare of antibiotics resistance related genes in the S. aureus isolated from milk of Arabian shecamels (26, 27), (28) demonstrate a small, soluble single-domain fragments derived from the unique variable region of dromedary heavy-chain antibodies which act as inhibitors to β-lactamase. this state arise from the camels rearing system in the desert regions of Iraq which leads to great decline in the therapeutic medication of the common bacterial infections. CHROMagar (CAS, MRSA) give best results for the detection of S. aureus and methicillin resistance pathogens comparatively with the conventional techniques, these results confirm data reported by (29). (30) has showed that primary streaking on (CAS) provides a convenient with less time consuming manner for the presumptive detection of S. aureus in routine clinical microbiology. (31) explained the very high accuracy of the use (CAS) medium and another author detected high sensitivity and specificity of the use (MRSA) medium (32). Conclusions, The high prevalence of antibiotic resistance suggests that subclinical mastitis in cattle plays a significant role in the spreading of multi-resistant staphylococci and represent a great public health concern in Iraq. Acknowledgments The author expresses his gratitude to the cattle and she-camels owners and the director of veterinary hospital in Al-Qadisyia for his cooperation in completing this study. The research has been carried out with the help of the Department of Animal Production in the College of Agriculture, University of Al-Qadisyia. References 1. Mohsenzadeh, M. & Fallah-Rad, A. H Prevalence of and antibiotic susceptibility of coagulase-negative Staphylococcus isolated from bovine intramammary infections in Mashhad. Iran. J. Ani. Vet. Adv., 6(8): Guliye, A. Y.; Van Creveld, C. & Yagil, R Detection of subclinical mastitis in dromedary camels (Camelus dromedarius) using somatic cell counts and the N- Acetyl-β-D-glucosaminidase test. Trop. Ani. Health Prod., 34: Hata, E.; Kobayashi, H.; Nakajima, H.; Shimizu, Y. & Eguchi, M. 2010a. Epidemiological analysis of Staphylococcus aureus isolated from cows and the environment of a dairy farm in Japan. J. Vet. Med. Sci., 72(5):

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